This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This project is part of a Roadmap Consortium (U54RR02437/ T. Woodruff, PI) entitled "The Oncofertility Consortium: Fertility Preservation for Women". It's goal is to validate options in nonhuman primates for restoring fertility in female cancer survivors by preventing ovarian exposure to the gametotoxic effects of therapy (by removing and preserving biopsies) and returning ovarian tissue, gametes or embryos. Studies in monkeys: (Aim 1) Bioengineer a scaffold that supports the three-dimensional architecture of the follicle and permits coordinated development of the follicle wall and oocyte in vitro;(Aim 2) Evaluate role of hormones and growth factors in promoting follicle growth and oocyte quality;(Aim 3) Optimize conditions for autotransplantation of ovarian cortex for coordinated follicle growth and oocyte maturation in vivo;and (Aim 4) Assess fertilization and embryonic potential of oocytes derived from in vitro matured follicles and transplanted ovarian cortical follicles in vivo. Immature follicles will be isolated from ovaries and cultured in an alginate matrix. Ovarian cortex will be autotransplanted to accessible sites (abdomen and forearm), and the ability of pro-angiogenic factors to restore ovarian function will be monitored. Mature oocytes produced by follicles in vitro and in cortical transplants will be evaluated for reproductive potential by in vitro fertilization, embryo transfer into surrogate mothers, pregnancy and health of offspring. Experiments were successful in growing small follicles for over 30 days in vitro to the antral stage, in promoting steroidogenic (progesterone, estrogen) and paracrine (anti-mullerian hormone, vascular endothelial growth factor) function, and producing meiotically mature eggs that fertilize in vitro.